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Biomedical subjects

J D Stanger

Publications and source records attributed to J D Stanger.

At least 19 recordsLinked to original sources
PubMed2001

Pregnancy following fertilization of zona-free, coronal cell intact human ova: Case Report.

A pregnancy by intracytoplasmic sperm injection (ICSI) of a couple whose oocytes were recovered without a zona pellucida is reported. Previous IVF/ICSI cycles indicated all oocytes had either very thin or no zona pellucida at the time of cumulus removal prior to ICSI. As a result, many ova were very fragile and lysed upon handling. In the latest attempt, six ova were recovered. After coronal cell dissection of two ova where both lacked a zona pellucida and one lysed, it was elected to attempt to inject the remaining four ova without removal of their coronal cells. The zona pellucida was not visible through the cell layer and the position of the polar body in three ova was speculative. Two embryos possessed normal pronuclei on day 1, and displayed six blastomeres prior to transfer on day 3. During preparation for transfer, sufficient coronal cells fell away to confirm both embryos lacked a zona pellucida. Pregnancy was confirmed by ultrasound observation of a fetal heart 4 weeks later. This report documents that where the zona pellucida is absent, the coronal cells provide sufficient support to maintain blastomere interaction and embryo viability and the zona pellucida is not essential for early embryo development and pregnancy.

Adult↗
PubMed1996

Pregnancy resulting from intracytoplasmic injection of cryopreserved spermatozoa recovered from testicular biopsy.

We report a clinical pregnancy occurring in a 31 year old patient following intracytoplasmic sperm injection (ICSI) of cryopreserved spermatozoa obtained from a testicular biopsy. This was the couple's second attempt at an ovarian stimulated cycle resulting in the collection of 17 metaphase II ova which were all injected with progressively motile spermatozoa. A fertilization rate of 58% and a cleavage rate of 90% were achieved. This report is our first case of ICSI using cryopreserved testicular spermatozoa which resulted in normal fertilization, embryo development and an on-going singleton pregnancy.

Adult↗
PubMed1992

Prefertilization treatment of mice with platelet activating factor affects pregnancy.

Embryo-derived platelet activating factor (EPAF) is thought to be either biologically similar to platelet activating factor (PAF) or responsible for PAF liberation in vivo. We have previously shown that premating PAF treatment in the mouse renders the platelets nonresponsive to EPAF, leading to a reduced implantation rate in these animals. In this study, we have shown that females, injected with PAF before mating, show altered embryo development invivo on day 4 postfertilization. This is manifested as an interruption of compaction, a reduced cell number per embryo, and reduced embryo number per mouse. Results suggest that EPAF represents an early pregnancy signal that supports embryo development. The most likely mechanism is via platelet activation, since only those mice that showed thrombocytopenia after fertilization were found to have normal embryos on day 4 postmating.

Animals↗
PubMed1991

Studies on murine embryo-derived platelet-activating factor (EPAF).

Studies were carried out using the splenectomized mouse bioassay (SMB) to investigate the nature of embryo-derived platelet-activating factor (EPAF) and its relationship to synthetic platelet activating factor (PAF). While both C16-PAF and embryo conditioned media (ECM) induced a significant platelet decline in the SMB at 15 min postinjection, C18-PAF induced a similar effect at 30 min postinjection. The degree of EPAF activity in ECM was not altered with increasing embryo number from 2 to 40/ml of media. In contrast, PAF (C16/C18 mixture) induced a linear increase in activity with increasing concentration, leading to lethal effects at high concentrations. While EPAF activity was not significantly altered when ECM was diluted 1/1,000, PAF activity was abolished at 1/10 dilution. EPAF in ECM was not inactivated by mouse plasma; however, lipid extracted ECM, like PAF, underwent rapid inactivation in the presence of plasma. Aggregometer studies using horse platelets showed that ECM and lipid-extracted ECM were unable to induce platelet aggregation, while thin-layer chromatography (TLC) purified ECM (Rf 0.23) successfully aggregated horse platelets in vitro. Results suggested that EPAF and PAF are not homologous. EPAF might consist of PAF bound to a regulatory carrier molecule and appears to be associated with EPAF-inhibitor substance(s) in ECM.

Animals↗
PubMed1988

Methods of water purification for the preparation of culture media in an IVF-ET programme.

Five methods of water purification for culture media preparation have been described. The quality of media thus prepared has been assessed retrospectively by relating to the fertilization and pregnancy rates of a total of 646 treatment cycles in an in-vitro fertilization and embryo transfer (IVF-ET) programme. All media for human use was subjected to quality control assessment in a mouse embryo culture system. Although no differences were seen in the fertilization rates or the proportion of women proceeding to embryo transfer, significant differences were seen in the pregnancy rates. The highest pregnancy rates were obtained using rainwater distilled six times (22/100, 22%) or eight times (40/184, 21.7%), and water polished using a Milli-Q water purifying system (41/181, 22.7%). Significantly lower pregnancy rates were obtained when the water was purified using an alternative Milli-Q system (9/89, 10.1%) or dam water distilled six times (6/63, 9.5%). These findings may reflect contamination of local dam water supplies but demonstrate the difficulty in purifying water for the preparation of culture medium.

Culture Media↗
PubMed1987

An evaluation of peripheral blood platelet enumeration as a monitor of fertilization and early pregnancy.

This paper reports our data that confirm the existence of early pregnancy-associated thrombocytopenia (EPAT) in the mouse and illustrate that the phenomenon is independent of age, parity, and strain differences. This paper also provides evidence that the EPAT phenomenon is induced by a soluble factor (EPAT-factor) released by the fertilized ovum. EPAT-factor was produced in vitro by mouse embryos from the 1-cell stage to the expanded blastocyst stage. The human study involved a "blind" analysis of serum samples, collected from in vitro fertilization-treated patients, for the presence of thrombocytopenic activity. Results suggest that measurement of this thrombocytopenic activity might be useful as an index of embryo viability and might be clinically applicable for the monitoring of implantation success in in vitro fertilization programs.

Animals↗
PubMed1987

Mechanistic studies of early pregnancy associated thrombocytopenia (EPAT) in the mouse.

The preimplantation period of uterine pregnancy is associated with the transient (first 4 days of gestation) expression of a state of early pregnancy-associated thrombocytopenia (EPAT), a phenomenon shown to be mediated by the embryo-derived EPAT factor, which presumably causes platelet activation and subsequent removal. We previously investigated the time course of production of EPAT factor in mouse embryo culture medium and found a correlation between the production of this factor and the in vivo platelet alterations in pregnant mice. The present paper supports the postulation that the EPAT factor and PAF-acether (a phospholipid platelet-activating factor) are related by providing data showing that PAF-acether may be responsible for the thrombocytopenia. Finally, data are presented to suggest that platelet activation, though not affecting the rate of ovulation, is important for successful ongoing pregnancy. Results suggest that the EPAT factor, produced by the fertilized egg, might act to signal uterine decidualization and/or modulate maternal immunological rejection of the implanting conceptus.

Animals↗
PubMed1985

Fetal abnormality (Goldenhar syndrome) occurring in one of triplet infants derived from in vitro fertilization with possible monozygotic twinning.

Of 36 infants delivered following conception by in vitro fertilization (IVF), 1 case of significant fetal abnormality has been detected. The infant is one of male triplets exposed to medroxyprogesterone acetate (MPA) in utero and demonstrates abnormalities of the ear, vertebral column, and ribs which fit clearly into the oculoauriculovertebral syndrome described by Goldenhar and which have been reported in monozygotic twins with discordance. Marker studies including eight blood groups, HLA haplotype assignments, and banding studies of the chromosomes indicate that the infant and one of his brothers may well be identical (P less than 0.001 for dizygosity) except for the Fy(a) antigen. It is deduced that the fetal abnormality is not causally related either to MPA exposure or to the techniques applied during fertilization and early embryo culture, and furthermore, if the infant is the monozygotic twin of one of his brothers, the two phenomena are unrelated, as each was likely to have occurred at different stages of embryonic development.

Abnormalities, Drug-Induced↗
PubMed1985

Hormonal profiles in the follicular phase, luteal phase and first trimester of pregnancies arising from in-vitro fertilization.

The hormonal profiles for oestradiol-17 beta, progesterone, prolactin and beta-human chorionic gonadotrophin (beta-hCG) are documented for the first 24 pregnancies arising from in-vitro fertilization during a collaborative project between the University of Western Australia and PIVET Laboratory. All patients had ovarian follicle stimulation with clomiphene citrate, sometimes combined with human menopausal gonadotrophin and all had oocyte recovery undertaken 36 h after injection of 5000 i.u. hCG. The follicular phase profile indicated that patients were admitted for the hCG injection when oestradiol-17 beta levels were around 1500 pmol/l per follicle with a dimension of greater than or equal to 1.6 cm on ultrasound. Luteal phase data indicated that oestradiol-17 beta and progesterone levels were two to three times higher than that expected during spontaneous conception cycles and those pregnancies which subsequently aborted had significantly lower levels in the late luteal phase. During pregnancy elevated oestradiol-17 beta and progesterone levels were maintained through the early weeks during organogenesis while the beta-hCG profile was similar to that reported for spontaneous pregnancies arising without ovarian stimulation. Six women aborted and the other 18 pregnancies have generated 22 infants.

Chorionic Gonadotropin↗
PubMed1985

Reduced in-vitro fertilization of human oocytes from patients with raised basal luteinizing hormone levels during the follicular phase.

A series of 62 women were managed in the University of Western Australia/PIVET Laboratory in-vitro fertilization programme. In 60 of them follicle growth was stimulated with clomiphene citrate with or without additional human menopausal gonadotrophin (hMG) and in two with hMG alone. Follicles were aspirated at laparoscopy following an hCG trigger injection and occasionally following a spontaneous luteinizing hormone (LH) surge. Oocytes were inseminated with 0.5 X 10(5)-10(5) sperm/ml 3-6 h later. A significant reduction (P less than 0.001) in the fertilization rate of mature oocytes was observed in those patients whose basal serum LH values were greater than 1 SD above the mean. Fifty-nine women subsequently had embryo transfer and of 10 clinical pregnancies, none occurred in those with elevated LH values. Reduced fertilization may be a reflection of premature oocyte maturation or ageing. This may have clinical implications in the management of some patients with unexplained infertility.

Clomiphene↗
PubMed1985

Relation between pregnancy-associated plasma protein A (PAPP-A) in human peri-ovulatory follicle fluid and the collection and fertilization of human ova in vitro.

In 42 patients, blood-free peri-ovulatory follicular fluid was aspirated from 86 follicles for estimation of pregnancy-associated plasma protein A (PAPP-A) and related to several variables associated with the fertilization of human ova in vitro. Follicle growth was stimulated by clomiphene citrate alone or in combination with human menopausal gonadotrophin (hMG) and ovulation was triggered by human chorionic gonadotrophin (hCG) except in six patients who had a spontaneous surge of luteinizing hormone (LH). PAPP-A was detected in 83 of the 86 follicles aspirated, ranging from 'undetectable' to 1044 micrograms/l. The concentration of PAPP-A was statistically significantly correlated with increasing follicle size, follicular oestradiol-17 beta concentration and oocyte recovery. At PAPP-A concentrations of greater than 200 micrograms/l, there was no correlation noted with the ability of the ovum to be fertilized, to cleave, or between PAPP-A levels in follicles from pregnant and non-pregnant cycles. There was no significant difference in the mean PAPP-A concentration for the two stimulation regimens, but concentrations were greater in follicular fluid from patients who surged spontaneously (P less than 0.001). It appears PAPP-A levels may provide an index of follicle maturity but not of the pregnancy potential of the ovum.

Body Fluids↗
PubMed1985

Antispermatozoal antibodies in human follicular fluid.

The possibility of antispermatozoal antibodies in women having significant effects in the higher regions of the female reproductive tract has been investigated. Follicular fluids (FF) and sera taken at the time of oocyte recovery from women undergoing in vitro fertilisation and embryo transfer (IVF-ET) were tested for the presence of antispermatozoal antibodies, and the concentrations of IgM, IgG, IgA, and complement C3 were determined. The concentrations of immunoglobulins and C3 in FF were consistent with transudation from serum inversely proportional to molecular weight. Titres of agglutinating and immobilizing antibodies in FF were usually one or two dilution steps below those of serum except where immobilizing activity was associated with IgM. IgG:IgA ratios were lower in FF from women with antispermatozoal antibodies, suggesting local production or enhanced transudation of IgA; however, a secretory component could not be detected in any of the follicular fluids in this study. Two women with antispermatozoal antibodies and infertility in excess of 5 years had successful IVF-ET and have delivered healthy infants.

Antibody Specificity↗
PubMed1985

The management of oligospermic infertility by in vitro fertilization.

The fertilization rates of mature preovulatory oocytes aspirated from 156 women treated by in vitro fertilization were analyzed as a function of spermatozoal density and motility and the findings were correlated with the category of infertility (chronic oligospermia, tubal disease, endometriosis, serum antibodies to spermatozoa in the female, and unexplained infertility). Overall reduced fertilization rates were found in all cases if the semen sample presented on the day of fertilization demonstrated less than or equal to 5 million motile spermatozoa per milliliter, less than 40% motile forms, or the combined findings of less than 20 million per ml and less than or equal to 60% motile forms. Where the husband was known to have chronic oligospermia, reduced fertilization was found if the semen on the day of fertilization contained less than 20 million spermatozoa per ml and less than 12 million motile spermatozoa per milliliter. For cases of unexplained infertility, a poor fertilization rate was noted if the semen demonstrated less than 60% progressively motile forms regardless of the overall spermatozoal density, implying that a proportion of unexplained infertility is due to a disorder of spermatozoa reflected by reduced motility. Pregnancies were achieved in 5 of 26 cases with chronic oligospermia, including 2 where oligospermia was very pronounced (less than or equal to 5 million motile spermatozoa per milliliter).

Endometriosis↗
PubMed1984

In-vitro fertilization in Western Australia. A viable service programme.

A service programme of in-vitro fertilization ( IVF ) and embryo transfer (ET) has been established in a small unit in Western Australia after a successful pilot study which was undertaken in 1981. The key features of the programme include simplified monitoring of the follicular phase of stimulated cycles; oocyte retrieval and ET are both undertaken during routine daytime work schedules. The results of the first eight months of the programme are presented, during which 13 further pregnancies were generated and 10 healthy infants were delivered (seven boys and three girls). In the last session in which 49 patients took part a mean of 2.3 mature preovulatory oocytes were collected by means of a double-lumen aspiration/flushing needle from 98% of 48 patients who reached the laparoscopy stage. Ninety-four per cent of 48 patients proceeded to embryo transfer by means of a double-catheter technique; the pregnancy rate was 20.8% per laparoscopy or 22.7% per embryo transfer.

Australia↗
PubMed1984

In-vitro fertilisation of oocytes from women with serum antisperm antibodies.

Five women whose infertility was believed to be related primarily to their serum antisperm antibodies underwent in-vitro fertilisation and embryo transfer. Follicle growth was stimulated with clomiphene citrate, sometimes combined with human menopausal gonadotropin, and ovulation was triggered off with human chorionic gonadotropin. 20 oocytes were collected from the five patients, and 15 of these were fertilised in the presence of donor serum. All embryos developed to morphologically normal 2-cell and 4-cell embryos. All five women proceeded to embryo transfer, and two became pregnant.

Antibodies↗
PubMed1984

The limitations of in vitro fertilization from males with severe oligospermia and abnormal sperm morphology.

Thirty-one patients whose infertility was attributed to oligospermia were included for treatment by in vitro fertilization and embryo transfer. Three subgroups were defined: severe oligospermia (less than or equal to 5 million motile sperm/ml), moderate oligospermia (6 to less than 12 million motile sperm/ml), and abnormal sperm morphology (greater than 60% atypical). The fertilization rates were compared to those of a normospermic group managed concurrently. A modified overlay technique of sperm preparation is described for oligospermic samples so that the number of motile spermatozoa inseminated into each tube or culture dish containing a mature preovulatory oocyte was similar in each category, within the range 0.5 to 2 X 10(5)/ml. Significantly fewer oocytes were fertilized in the severe oligospermic group (P less than 0.001), suggesting a reduced capacity for fertilization by spermatozoa from severely oligospermic males. The fertilization rate of oocytes was normal in the moderate oligospermic group and those with abnormal morphology, although in the latter there was a significant delay noted in reaching the pronuclear stage (P less than 0.001), and the embryos were at a less advanced stage of cleavage at the time of transfer (0.001 less than P less than 0.01). Pregnancies were achieved in both the severe and the moderate oligospermic groups, with healthy infants delivered from each.

Embryo Transfer↗